NDRG2调控HGF/c-MET通路抑制结肠癌细胞的增殖

NDRG2 regulates HGF/c-MET signaling pathway to inhibit the proliferation of colon cancer cells

目的:探讨抑癌基因NDRG2通过调节HGF/c-MET信号通路,抑制结肠癌HT29细胞增殖的机制。方法:建立NDRG2过表达细胞株HT29-NDRG2及对照组HT29-Cherry,并于第1、2、3、4、5天以MTT法检测细胞增殖;以不同浓度的HGF(0ng/ml、1ng/ml、2ng/ml、4ng/ml、8ng/ml、10ng/ml)刺激HT29亲本细胞,并于第1、2、3天以MTT法检测HT29细胞的增殖;采用qPCR法检测NDRG2过表达细胞株HT29-NDRG2与对照组HT29-Cherry两组细胞中HGF的表达水平;Western blot检测HT29-Cherry和HT29-NDRG2两组细胞以及添加HGF(10ng/ml)刺激HT29亲本细胞后各组细胞中HGF/c-MET通路中关键分子的表达。结果:过表达NDRG2以及HGF刺激均可以显著抑制HT29细胞的增殖能力;NDRG2的高表达可以上调HGF的转录水平;HGF可以刺激c-MET和p-ERK1/2,并上调p21和p27,抑制HT29细胞的增殖;过表达NDRG2明显上调c-MET、p-ERK1/2,并诱导p 21和p 27的高表达,从而抑制HT29细胞的增殖。结论:NDRG2可以通过调节HGF/c-MET信号通路从而抑制结肠癌细胞HT29细胞的增殖能力。

Objective：To investigate whether tumor suppressor gene NDRG2 could inhibitthe proliferation of the colon cancer cells HT29 by regulating HGF/c MET signaling pathway. Methods： Construct stable cell lines of HT29 NDRG2 with overexpression of NDRG2 and the control group of HT29 Cherry. HT29 Cherry and HT29 NDRG2 cells, and parental HT29 cells were planted into 96 well plates, HT29 Cherry and HT29 NDRG2 cells were cultured normally, and the proliferation rates were detected by MTr analysis at 1,2,3,4,5 days. The parental HT29 cells were treated with different concentration of HGF （ 0ng/ml, 1 ng/ml, 2ng/m1,4ng/ml, 8ng/ml, 10ng/ml） for 1,2,3 days and identified the proliferation rates by MTr analysis. The relative expression of HGF in HT29 Cherry and HT29 NDRG2 cell lines were verified by qPCR. HT29 Cherry and HT29 NDRG2 cells were prepared nor mally, and treated the parental HT29 cells with HGF （10ng/ml） for 0,12,24h. Western blot was employed to detect the expression of pivotal molecules in HGF/c MET signaling pathway. Results ：The proliferation rates of HT29 cells were obviously suppressed with overexpression of NDRG2, as well as when treated with dose dependent HGF. Over expression of NDRG2 could enhance the transcriptional level of HGF. HGF stimulation and overexpression of NDRG2 would up regulate c MET and p ERK1/2,thus enhance p21 and p27, and suppresse the proliferation rates of HT29 cells. Conclusion ： NDRG2 may regulate HGF/c MET signaling pathway to inhibitproliferation rate of HT29 cells.